Compositions and methods for the prevention and diagnosis of Lyme disease

Abstract

Methods and compositions for the prevention and diagnosis of Lyme disease. OspA and OspB polypeptides and serotypic variants thereof, which elicit in a treated animal the formation of an immune response which is effective to treat or protect against Lyme disease as caused by infection with B. burgdorferi. Anti-OspA and anti-OspB antibodies that are effective to treat or protect against Lyme disease as caused by infection with B. burgdorferi. A screening method for the selection of those OspA and OspB polypeptides and anti-OspA and anti-OspB antibodies that are useful for the prevention and detection of Lyre disease. Diagnostic kits including OspA and OspB polypeptides or antibodies directed against such polypeptides.

We claim:  
     
       1. An isolated polypeptide comprising at least 50 contiguous amino acids of SEQ ID NO: 10, wherein the polypeptide is immunogenic. 
     
     
       2. A fusion protein comprising a first polypeptide and an additional polypeptide, wherein said first polypeptide is a polypeptide according to claim  1 . 
     
     
       3. A vaccine comprising a fusion protein according to claim  2 . 
     
     
       4. A multimeric protein comprising a polypeptide according to claim  1 . 
     
     
       5. A vaccine comprising a multimeric protein according to claim  4 . 
     
     
       6. A vaccine comprising a polypeptide according to claim  1 . 
     
     
       7. An isolated polypeptide selected from the group consisting of: 
       (a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 10; and  
       (b) a polypeptide comprising the amino acid sequence from position 2 to 273 of SEQ ID NO: 10.  
     
     
       8. The polypeptide according to claim  7 , wherein the polypeptide contains one or more modifications selected from the group consisting of: reaction of free amino, carboxyl or hydroxyl side groups of the amino acid residues of the polypeptide, and reaction with acids or bases to form salts of the protein or polypeptide, wherein said polypeptide containing said modifications retains the immunogenicity of the unmodified polypeptide. 
     
     
       9. A diagnostic kit comprising instructions for use and a polypeptide according to any one of claims  7 ,  8  or  1 . 
     
     
       10. A fusion protein comprising a first polypeptide and an additional polypeptide, wherein said first polypeptide is a polypeptide according to claim  8 . 
     
     
       11. A vaccine comprising a fusion protein according to claim  10 . 
     
     
       12. A multimeric protein comprising a polypeptide according to claim  8 . 
     
     
       13. A vaccine comprising a multimeric protein according to claim  12 . 
     
     
       14. A vaccine comprising a polypeptide according to claim  8 . 
     
     
       15. A fusion protein comprising a first polypeptide and an additional polypeptide, wherein said first polypeptide is a polypeptide according to claim  7 . 
     
     
       16. A diagnostic kit comprising instructions for use and a fusion protein according to any one of claims  15 ,  10  or  2 . 
     
     
       17. A vaccine comprising a fusion protein according to claim  15 . 
     
     
       18. The vaccine according to any one of claims  17 ,  11  or  3 , wherein the fusion protein is crosslinked to an immunogenic carrier. 
     
     
       19. The vaccine according to claim  18 , further comprising an adjuvant. 
     
     
       20. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  19 . 
     
     
       21. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  18 . 
     
     
       22. The vaccine according to any one of claims  17 ,  11  or  3 , further comprising an adjuvant. 
     
     
       23. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  22 . 
     
     
       24. A vaccine comprising a polypeptide according to claim  7 . 
     
     
       25. The vaccine according to any one of claims  24 ,  14  or  6 , wherein the polypeptide is crosslinked to an immunogenic carrier. 
     
     
       26. The vaccine according to claim  25 , further comprising an adjuvant. 
     
     
       27. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a multimeric protein according to claim  26 . 
     
     
       28. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a fusion protein according to claim  25 . 
     
     
       29. The vaccine according to any one of claims  24 ,  14  or  6 , further comprising an adjuvant. 
     
     
       30. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  29 . 
     
     
       31. A multimeric protein comprising a polypeptide according to claim  7 . 
     
     
       32. A diagnostic kit comprising instructions for use and a multimeric protein according to any one of claims  31 ,  12  or  4 . 
     
     
       33. A vaccine comprising a multimeric protein according to claim  31 . 
     
     
       34. The vaccine according to any one of claims  33 ,  13  or  5 , wherein the multimeric protein is crosslinked to an imnmunogenic carrier. 
     
     
       35. The vaccine according to claim  34 , further comprising an adjuvant. 
     
     
       36. A method for treating or preventing  B. Burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  35 . 
     
     
       37. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  34 . 
     
     
       38. The vaccine according to any one of claims  33 ,  13  or  5 , further comprising an adjuvant. 
     
     
       39. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  38 . 
     
     
       40. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a composition selected from the group consisting of: 
       (a) a polypeptide according to any one of claims  7 ,  8  or  1 ;  
       (b) a fusion protein according to any one of claims  15 ,  10  or  2 ;  
       (c) a multimeric protein according to any one of claims  31 ,  12  or  4 ; and  
       (d) a vaccine according to any one of claims  3 ,  5 ,  6 ,  11 ,  13 ,  14 ,  17 ,  24 , or  33 .  
     
     
       41. A method for detecting  B. burgdorferi  infection comprising the step of assaying a body fluid of a suspected infected mammalian host with a polypeptide according to any one of claims  7 ,  8  or  1 , wherein said body fluid is contacted with said polypeptide and a complex comprising said polypeptide and an antibody in said body fluid is detected, said complex being indicative of  B. burgdorferi  infection. 
     
     
       42. A method for detecting  B. burgdorferi  infection comprising the step of assaying a body fluid of a suspected infected mammalian host with a fusion protein according to any one of claims  15 ,  10  or  2 , wherein said body fluid is contacted with said fusion protein and a complex comprising said fusion protein and an antibody in said body fluid is detected, said complex being indicative of  B. burgdorferd  infection. 
     
     
       43. A method for detecting  B. burgdorferi  infection comprising the step of assaying a body fluid of a suspected infected mammalian host with a multimeric protein according to any one of claims  31 ,  12  or  4  wherein said body fluid is contacted with said multimeric protein and a complex comprising said multimeric protein and an antibody in said body fluid is detected, said complex being indicative of  B. burgdorferi  infection. 
     
     
       44. An isolated polypeptide comprising amino acids 149-164 of SEQ ID NO: 4, wherein the polypeptide is immunogenic. 
     
     
       45. A fusion protein comprising a first polypeptide and an additional polypeptide, wherein said first polypeptide is a polypeptide according to claim  44 . 
     
     
       46. A vaccine comprising a fusion protein according to claim  45 . 
     
     
       47. A multimeric protein comprising a polypeptide according to claim  44 . 
     
     
       48. A vaccine comprising a multimeric protein according to claim  47 . 
     
     
       49. A vaccine comprising a polypeptide according to claim  44 . 
     
     
       50. An isolated polypeptide selected from the group consisting of: 
       (a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 4;  
       (b) a polypeptide comprising the amino acid sequence from position 2 to 273 of SEQ ID NO: 4;  
       (c) a polypeptide comprising the amino acid sequence encoded by nucleotides 200-819 of SEQ ID NO: 3; and  
       (d) a polypeptide comprising the amino acid sequence encoded by nucleotides 400-819 of SEQ ID NO: 3.  
     
     
       51. The isolated polypeptide according to claim  50 , wherein the polypeptide contains one or more modifications selected from the group consisting of: reaction of free amino, carboxyl or hydroxyl side groups of the amino acid residues of polypeptide, and reaction with acids or bases to form salts of the protein or polypeptide, wherein the polypeptide containing the modifications retains the immunogenicity of the unmodified polypeptide. 
     
     
       52. A fusion protein comprising a first polypeptide and an additional polypeptide, wherein said first polypeptide is a polypeptide according to claim  51 . 
     
     
       53. A vaccine comprising a fusion protein according to claim  52 . 
     
     
       54. A multimeric protein comprising a polypeptide according to claim  51 . 
     
     
       55. A vaccine comprising a multimeric protein according to claim  54 . 
     
     
       56. A vaccine comprising a polypeptide according to claim  51 . 
     
     
       57. A diagnostic kit comprising instructions for use and a polypeptide according to any one of claims  50 ,  51  or  44 . 
     
     
       58. A fusion protein comprising a first polypeptide and an additional polypeptide, wherein said first polypeptide is a polypeptide according to claim  50 . 
     
     
       59. A diagnostic kit comprising instructions for use and a fusion protein according to any one of claims  58 ,  53  or  46 . 
     
     
       60. A vaccine comprising a fusion protein according to claim  58 . 
     
     
       61. The vaccine according to claim  60 , wherein the fusion protein is crosslinked to an immunogenic carrier. 
     
     
       62. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  61 . 
     
     
       63. The vaccine according to claim  61 , further comprising an adjuvant. 
     
     
       64. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  63 . 
     
     
       65. The vaccine according to claim  60 , further comprising an adjuvant. 
     
     
       66. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  65 . 
     
     
       67. A multimeric protein comprising a polypeptide according to claim  50 . 
     
     
       68. A diagnostic kit comprising instructions for use and a multimeric protein according to any one of claims  67 ,  55  or  48 . 
     
     
       69. A vaccine comprising a multimeric protein according to claim  67 . 
     
     
       70. The vaccine according to claim  69 , wherein the multimeric protein is crosslinked to an immunogenic carrier. 
     
     
       71. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  70 . 
     
     
       72. The vaccine according to claim  70 , further comprising an adjuvant. 
     
     
       73. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  72 . 
     
     
       74. The vaccine according to claim  69 , further comprising an adjuvant. 
     
     
       75. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  74 . 
     
     
       76. A vaccine comprising a polypeptide according to claim  50 . 
     
     
       77. The vaccine according to claim  76 , wherein the polypeptide is crossing to an immunogenic carrier. 
     
     
       78. The vaccine according to claim  77 , further comprising an adjuvant. 
     
     
       79. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a multimeric protein according to claim  78 . 
     
     
       80. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a fusion protein according to claim  77 . 
     
     
       81. The vaccine according to claim  76 , further comprising an adjuvant. 
     
     
       82. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a vaccine according to claim  81 . 
     
     
       83. A method for treating or preventing  B. burgdorferi  infection and Lyme disease comprising the step of administering to a patient a composition selected from the group consisting of: 
       (a) a polypeptide according to any one of claims  50 ,  51  or  49 ;  
       (b) a fusion protein according to any one of claims  58 ,  53  or  46 ;  
       (c) a multimeric protein according to any one of claims  67 ,  55  or  48 ; and  
       (d) a vaccine according to any one of claims  76 ,  60 , or  69 .  
     
     
       84. A method for detecting  B. burgdorferi  infection comprising the step of assaying a body fluid of a suspected infected mammalian host with a polypeptide according to any one of claims  50 ,  51  or  44 , wherein said body fluid is contacted with said polypeptide and a complex comprising said polypeptide and an antibody in said body fluid is detected, said complex being indicative of  B. burgdorferi  infection. 
     
     
       85. A method for detecting  B. burgdorferi  infection comprising the step of assaying a body fluid of a suspected infected mammalian host with a fusion protein according to any one of claims  58 ,  53  or  46 , wherein said body fluid is contacted with said fusion protein and a complex comprising said fusion protein and an antibody in said body fluid is detected, said complex being indicative of  B. burgdorferi  infection. 
     
     
       86. A method for detecting  B. burgdorferi  infection comprising the step of assaying a body fluid of a suspected infected mammalian host with a multimeric protein according to any one of claims  67 ,  55  or  48 , wherein said body fluid is contacted with said multimeric protein and a complex comprising said multimeric protein and an antibody in said body fluid is detected, said complex being indicative of  B. burgdorferi  infection.